Biol Reprod Keystone Symposia Conference on Frontiers in Reproductive Biology & Regulation of Fertility.
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BOR - Papers in Press, published online ahead of print September 15, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.032078
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Submitted May 13, 2004
Returned for revision June 8, 2004
Accepted August 23, 2004

Mechanisms of Hormone Action


Nitric Oxide Mediates Inhibitory Effect of Leptin on Insulin-Like Growth Factor I Augmentation of 17{beta}-Estradiol Production in Human Granulosa Cells

He-Feng Huang , Bo Wang , Xiao-Fu Yang , Qiong Luo , and Jian-Zhong Sheng *

* To whom correspondence should be addressed. E-mail: jzsheng{at}ucalgary.ca.

Abstract
In the present study the authors investigated if the inhibitory effect of leptin in the ovary was mediated via nitric oxide (NO) using human granulosa cells (GCs). Human GCs were obtained from pre-ovulatory follicles of women underwent IVF. Reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that human GCs expressed mRNA of leptin and mRNA of isoforms of leptin receptor including one long form and two types of short forms. Exposure of human GCs to leptin at concentrations of 3 to 30 ng/ml for 60 min dose-dependently increased the fluorescence of 4,5-diaminofluorescein (DAF-2), an NO-sensitive dye. The effect of leptin on DAF-2 fluorescence was inhibited by pretreatment of human GCs with 100 µM nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthase (NOS), indicating that the increase in DAF-2 fluorescence properly reflected the intracellular NO production. FSH (1 ng/ml ) and IGF-I (30 ng/ml) stimulated 17{beta}-estradiol (E2) production in human GCs, respectively. FSH plus IGF-I induced a further increase in E2 production. Leptin did not significantly alter basal or FSH-dependent E2 production, but it inhibited the effect of IGF-I on E2 production and the synergistic effect of IGF-I on FSH-stimulated E2 production. The inhibitory effect of leptin on IGF-I argumentation of E2 production was attenuated by pretreatment of human GCs with 100 µM L-NAME. In conclusion, leptin could induce NO production in human GCs. The inhibitory effect of leptin on IGF-I augmentation of E2 production in human GCs was attenuated by L-NAME, strongly suggesting that NO may mediate the action of leptin in human GCs.

Key words: Estradiol • Granulosa cells • Leptin • Leptin receptor • Nitric oxide


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A G Ricci, M P Di Yorio, and A G Faletti
Inhibitory effect of leptin on the rat ovary during the ovulatory process.
Reproduction, November 1, 2006; 132(5): 771 - 780.
[Abstract] [Full Text] [PDF]


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